SmartCells™ Chemically Competent E. coli BioVector NTCC质粒载体菌种细胞基因保藏中心

SmartCells™ Chemically Competent E. coli

Description: gWIZ vectors represent
a new series of plasmids that have been
engineered to produce the highest levels
of transgene expression in a wide range of
mammalian cells and tissues. It contains a
proprietarily modified promoter followed
by the intron A from the human
cytomegalovirus (CMV) immediate early
gene and a high-efficiency artificial
transcription terminator. The expression
vector is constructed in the context of a
plasmid backbone extensively modified to
achieve the enhanced levels of trangene
expression in mammalian cells as well as
high efficiency of plasmid production in
E. coli.
Components: 25 µg gWIZ GFP plasmid in
25 µl sterile TE buffer.
Storage: Store at -20o
C.
Comments: gWIZ is suitable for in vitro and in vivo
gene expression studies and applications.
Use Kanamycin as selection to grow the
plasmid in E. coli.
INTRODUCTION
The CMV immediate early gene (IE) promoter/enhancer is
the most widely used constitutive promoter for expressing
high levels of trangene product in many mammalian cells
and tissues. However, not all CMV IE gene
promoter/enhancer-based expression vectors are created
equal. Depending on the actual CMV IE gene sequences
used and the context of the plasmid backbone upon which
the expression cassette is constructed, the expression levels
can vary as much as two orders of magnitude. The CMV IE
promoter sequences contained in the gWIZ vectors have
been systemically analyzed and modified. The
modifications include removing the sequences that are
redundant and deleterious to the high levels of expression
while retaining those sequences that are of high
transcriptional potency. After coupling the modified
promoter with a high-efficiency synthetic transcriptional
terminator, the whole expression cassette is finally
constructed on a plasmid backbone that has also been
streamlined and modified to accommodate the high levels of
expression in mammalian cells as well as high yield of
plasmid production in E. coli. The resulting plasmid, gWIZ
expression vector, is capable of fully unleashing the
potential of the CMV promoter and giving the highest levels
of expression possible both in vitro and in vivo.
USAGE
• For extremely high levels of transgene expression in
mammalian cells and tissues
• Can be used with GenePORTER 2 (Cat. # T202007 or
T202015) to transfect a wide variety of mammalian cells
and tissues
DETECTION OF THE EXPRESSED GENE
Green Fluorescent Protein: For transfections conducted
with plasmids encoding GFP, the detection can be done by
epifluorescence or confocal microscopy. The GFP produced
has an excitation peak at 470-480 nm and emission peak at
510 nm. Expression level of GFP can also be monitored by
fluorescence-activated cell sorter analysis as described by L.
Cheng et al. * The gWIZ-GFP plasmid contains a modified
and much brighter version of green fluorescent protein gene.
*Cheng, L. et al. (1996) Use of green fluorescent protein variants to
monitor gene transfer and expression in mammalian cells. Nature
Biotechnology 14, 606-609.
RELATED PRODUCTS
Product Cat. Nos.
gWIZ Blank Vector P000200
gWIZ β-Gal Vector P010200
gWIZ CAT Vector P020200
gWIZ Luciferase Vector P030200
gWIZ Secreted AP Vector P050200
phCMV1 Vector Kit P003100
phCMV2 Vector Kit P003200
phCMV3 Vector Kit P003300
phCMV-GFP FSR Vector P003400
phCMV-Luciferase FSR Vector P003500
TurboCells® Chemically Competent E. coli
C300020
SmartCells™ Chemically Competent E. coli
C101020
GenePORTER® 2 Transfection Reagent
75 transfections (0.75 ml) T202007
150 transfections (1.5 ml) T202015

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